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09-20-2007, 12:03 AM
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 problem in centrifuge of protein hi I`m making 1- centrifuge the meat tissue hemogenate at 1400 rpm 2- then recentrifuge at 15000 rpm for 15 minutes under cooling 3-then put 40 micro of supernatent + 10 micro loading buffer 4-then denaturate at 95 c for 5 minute 5-then centrifuge at 15000 rpm under cooling every time I make this step but now I make the step number 2 but not under cooling then at the end of work the bands of protein is very weak so I`m asking is not cooling desruct the protein also I ask can I neglect the step number 5 that mean put the supernatent+loading buffer and denaturate only then inject it in the well please reply me quickly to repeat the work thanks        |
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